Agarose degradation for utilization: Enzymes, pathways, metabolic engineering methods and products

Red algae are important renewable bioresources with very large annual outputs. Agarose is the major carbohydrate component of many red algae and has potential to be of value in the production of agaro-oligosaccharides, biofuels and other chemicals. In this review, we summarize the degradation pathway of agarose, which includes an upstream part involving transformation of agarose into its two monomers, D-galactose (D-Gal) and 3,6-anhydro-α-L-galactose (L-AHG), and a downstream part involving monosaccharide degradation pathways. The upstream part involves agarolytic enzymes such as α-agarase, β-agarase, α-neoagarobiose hydrolase, and agarolytic β-galactosidase. The downstream part includes the degradation pathways of D-Gal and L-AHG. In addition, the production of functional agaro-oligosaccharides such as neoagarobiose and monosaccharides such as L-AHG with different agarolytic enzymes is reviewed. Third, techniques for the setup, regulation and optimization of agarose degradation to increase utilization efficiency of agarose are summarized. Although heterologous construction of the whole agarose degradation pathway in an engineered strain has not been reported, biotechnologies applied to improve D-Gal utilization efficiency and construct L-AHG catalytic routes are reviewed. Finally, critical aspects that may aid in the construction of engineered microorganisms that can fully utilize agarose to produce agaro-oligosaccharides or as carbon sources for production of biofuels or other value-adding chemicals are discussed.